[1]刘白朵,李风良,和碧蕾,等.溴氰菊酯胁迫下小菜蛾差异表达基因的筛选与分析[J].南京师大学报(自然科学版),2010,33(02):91-95.
　Liu Baiduo,Li Fengliang,He Bilei,et al.Screening and Analysis of the Differential Expressed Sequence of Diamondback Moth,Plutella xylostella[J].Journal of Nanjing Normal University(Natural Science Edition),2010,33(02):91-95.
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溴氰菊酯胁迫下小菜蛾差异表达基因的筛选与分析()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第33卷
期数:
2010年02期
页码:
91-95
栏目:
生命科学
出版日期:
2010-06-20

文章信息/Info

Title:
Screening and Analysis of the Differential Expressed Sequence of Diamondback Moth,Plutella xylostella
作者:
刘白朵1;  李风良2;  和碧蕾1;  于希忠1;  李忠英2;  程罗根1
1. 南京师范大学生命科学学院, 江苏南京210046 2. 贵州省农业科学院植物保护研究所, 贵州贵阳550006
Author(s):
Liu Baiduo1; Li Fengliang2; He Bilei1; Yu Xizhong1; Li Zhongying2; Cheng Luogen1
1.School of Life Science,Nanjing Normal University,Nanjing 210046,China 2. Institute of Plant P rotect ion, Gu izhou A cadem y of Agricu ltu re Sciences, Gu iyang 550006, China
关键词:
小菜蛾;  抗性遗传;  菌落PCR;  cDNA RDA
Keywords:
Plutella X y lostella ( L. );  inher itance of res istance;  co lony PCR;  cDNA RDA
分类号:
S433
摘要:
通过双向cDNA RDA,即分别以敏感品系为检测子(tester)、抗溴氰菊酯品系为驱赶子(driver)和以抗溴氰菊酯品系为检测子(tester)、敏感品系为驱赶子(driver)筛选2种品系中的差异表达基因,初步分析小菜蛾敏感品系和抗性品系之间的遗传差异.经过3轮消减杂交后,将所得的差异片段克隆于PMD 18-T载体,氨苄筛选阳性克隆,经菌落PCR鉴定后送样测序.得到2条序列与GenBank数据库中的部分已知序列有一定同源性,其中1条序列与编码S3a蛋白基因有较高的同源性.
Abstract:
W e sifted the d ifferential expressed sequence from P lutella xy los tella susceptible stra in and the deltam ethrinres istance stra in by tw o-w ay cDNA representa tiona l d ifference ana ly sis( cDNA RDA). The tw o-way cDNA RDA was perform ed respective ly using cDNA from the P. xy loste lla susceptib le stra in as Tester amp licon and from the de ltame thr in-res istance stra in as Dr iver am plicon, and us ing cDNA from the deltam ethrin-resistance stra in as Tester am plicon and from the susceptib le stra in as Dr iver amp licon. The differential sequences be tw een P lutella xy los tella susceptib le stra in and the de ltam ethrin- resistance stra in w ere analysed prelim ina rily. A fter three rounds o f subtractive hybr idiza tion, the d ifferentia l fragm entsw ere cloned w ith pMD18-T vector, then screen ing positive clone w ith amp ic illin and sequenced a fter co-l ony PCR. Compar isons between our sequencing result and da ta in GenB ank show ed that therew ere two sequences appearing re la tive ly high hom ology to some know n sequences. One o f them had h igh hom o logy w ith the sequence encoding s3a prote in.

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备注/Memo

备注/Memo:
基金项目: 国家自然科学基金( 30560085)、南京师范大学实践创新训练计划项目. 通讯联系人: 程罗根, 博士, 教授, 硕士生导师, 研究方向: 遗传学. E-mail:chengluogen@njnu. edu. cn
更新日期/Last Update: 2013-04-08